scholarly journals Correlative 3D microscopy of single cells using super-resolution and scanning ion-conductance microscopy

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Vytautas Navikas ◽  
Samuel M. Leitao ◽  
Kristin S. Grussmayer ◽  
Adrien Descloux ◽  
Barney Drake ◽  
...  
Keyword(s):  
High Resolution ◽  
Biological Samples ◽  
Single Cells ◽  
Super Resolution ◽  
Live Cell ◽  
Actin Dynamics ◽  
Label Free ◽  
Axial Resolution ◽  
Ion Conductance ◽  
Scanning Ion Conductance Microscopy

AbstractHigh-resolution live-cell imaging is necessary to study complex biological phenomena. Modern fluorescence microscopy methods are increasingly combined with complementary, label-free techniques to put the fluorescence information into the cellular context. The most common high-resolution imaging approaches used in combination with fluorescence imaging are electron microscopy and atomic-force microscopy (AFM), originally developed for solid-state material characterization. AFM routinely resolves atomic steps, however on soft biological samples, the forces between the tip and the sample deform the fragile membrane, thereby distorting the otherwise high axial resolution of the technique. Here we present scanning ion-conductance microscopy (SICM) as an alternative approach for topographical imaging of soft biological samples, preserving high axial resolution on cells. SICM is complemented with live-cell compatible super-resolution optical fluctuation imaging (SOFI). To demonstrate the capabilities of our method we show correlative 3D cellular maps with SOFI implementation in both 2D and 3D with self-blinking dyes for two-color high-order SOFI imaging. Finally, we employ correlative SICM/SOFI microscopy for visualizing actin dynamics in live COS-7 cells with subdiffraction-resolution.

scholarly journals Correlative 3D microscopy of single cells using super-resolution and scanning ion-conductance microscopy

2020 ◽  
Author(s):  
Vytautas Navikas ◽  
Samuel M. Leitao ◽  
Kristin S. Grussmayer ◽  
Adrien Descloux ◽  
Barney Drake ◽  
...  
Keyword(s):  
High Resolution ◽  
Biological Samples ◽  
Single Cells ◽  
Super Resolution ◽  
Live Cell ◽  
Actin Dynamics ◽  
Label Free ◽  
Axial Resolution ◽  
Ion Conductance ◽  
Scanning Ion Conductance Microscopy

AbstractHigh-resolution live-cell imaging is necessary to study complex biological phenomena. Modern fluorescence microscopy methods are increasingly combined with complementary, label-free techniques to put the fluorescence information into the cellular context. The most common high-resolution imaging approaches used in combination with fluorescence imaging are electron microscopy and atomic-force microscopy (AFM), originally developed for solid-state material characterization. AFM routinely resolves atomic steps, however on soft biological samples, the forces between the tip and the sample deform the fragile membrane, thereby distorting the otherwise high axial resolution of the technique. Here we present scanning ion-conductance microscopy (SICM) as an alternative approach for topographical imaging of soft biological samples, preserving high axial resolution on cells. SICM is complemented with live-cell compatible super-resolution optical fluctuation imaging (SOFI). To demonstrate the capabilities of our method we show correlative 3D cellular maps with SOFI implementation in both 2D and 3D with self-blinking dyes for two-color high-order SOFI imaging. Finally, we employ correlative SICM/SOFI microscopy for visualizing actin dynamics in live COS-7 cells with subdiffractional resolution.

scholarly journals 3D phonon microscopy with sub-micron axial-resolution

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Richard J. Smith ◽  
Fernando Pérez-Cota ◽  
Leonel Marques ◽  
Matt Clark
Keyword(s):  
Signal To Noise Ratio ◽  
Single Cells ◽  
Optical Resolution ◽  
Acquisition Time ◽  
Label Free ◽  
Axial Resolution ◽  
Force Microscopy ◽  
Atomic Force ◽  
Accuracy And Precision ◽  
Good Agreement

AbstractBrillouin light scattering (BLS) is an emerging method for cell imaging and characterisation. It allows elasticity-related contrast, optical resolution and label-free operation. Phonon microscopy detects BLS from laser generated coherent phonon fields to offer an attractive route for imaging since, at GHz frequencies, the phonon wavelength is sub-optical. Using phonon fields to image single cells is challenging as the signal to noise ratio and acquisition time are often poor. However, recent advances in the instrumentation have enabled imaging of fixed and living cells. This work presents the first experimental characterisation of phonon-based axial resolution provided by the response to a sharp edge. The obtained axial resolution is up to 10 times higher than that of the optical system used to take the measurements. Validation of the results are obtained with various polymer objects, which are in good agreement with those obtained using atomic force microscopy. Edge localisation, and hence profilometry, of a phantom boundary is measured with accuracy and precision of approximately 60 nm and 100 nm respectively. Finally, 3D imaging of fixed cells in culture medium is demonstrated.

Scanning Ion Conductance Microscopy for High-Resolution Topography of Soft Samples Including Live Cells

2011 ◽  
Vol 17 (S2) ◽  
pp. 236-237
Author(s):  
G De Filippi ◽  
C Moore
Keyword(s):  
High Resolution ◽  
Live Cells ◽  
Ion Conductance ◽  
Scanning Ion Conductance Microscopy ◽  
Soft Samples

Extended abstract of a paper presented at Microscopy and Microanalysis 2011 in Nashville, Tennessee, USA, August 7–August 11, 2011.

scholarly journals Super-Resolution Label-free Volumetric Vibrational Imaging

2021 ◽  
Author(s):  
Chenxi Qian ◽  
Kun Miao ◽  
Li-En Lin ◽  
Xinhong Chen ◽  
Jiajun Du ◽  
...  
Keyword(s):  
High Resolution ◽  
Three Dimensional ◽  
Super Resolution ◽  
Label Free ◽  
Fluorescence Techniques ◽  
Biological Structures ◽  
Imaging Strategy ◽  
Endogenous Proteins ◽  
Low Efficiency ◽  
Protein Rich

Innovations in high-resolution optical imaging have allowed visualization of nanoscale biological structures and connections. However, super-resolution fluorescence techniques, including both optics-oriented and sample-expansion based, are limited in quantification and throughput especially in tissues from photobleaching or quenching of the fluorophores, and low-efficiency or non-uniform delivery of the probes. Here, we report a general sample-expansion vibrational imaging strategy, termed VISTA, for scalable label-free high-resolution interrogations of protein-rich biological structures with resolution down to 82 nm. VISTA achieves decent three-dimensional image quality through optimal retention of endogenous proteins, isotropic sample expansion, and deprivation of scattering lipids. Free from probe-labeling associated issues, VISTA offers unbiased and high-throughput tissue investigations. With correlative VISTA and immunofluorescence, we further validated the imaging specificity of VISTA and trained an image-segmentation model for label-free multi-component and volumetric prediction of nucleus, blood vessels, neuronal cells and dendrites in complex mouse brain tissues. VISTA could hence open new avenues for versatile biomedical studies.

scholarly journals High-resolution label-free 3D mapping of extracellular pH of single living cells

2019 ◽  
Vol 10 (1) ◽  
Author(s):  
Yanjun Zhang ◽  
Yasufumi Takahashi ◽  
Sung Pil Hong ◽  
Fengjie Liu ◽  
Joanna Bednarska ◽  
...  
Keyword(s):  
Single Cell ◽  
Cancer Cells ◽  
Extracellular Ph ◽  
Single Cell Level ◽  
Ph Sensing ◽  
Label Free ◽  
Cell Level ◽  
Spatiotemporal Resolution ◽  
Ion Conductance ◽  
Scanning Ion Conductance Microscopy

AbstractDynamic mapping of extracellular pH (pHe) at the single-cell level is critical for understanding the role of H+ in cellular and subcellular processes, with particular importance in cancer. While several pHe sensing techniques have been developed, accessing this information at the single-cell level requires improvement in sensitivity, spatial and temporal resolution. We report on a zwitterionic label-free pH nanoprobe that addresses these long-standing challenges. The probe has a sensitivity > 0.01 units, 2 ms response time, and 50 nm spatial resolution. The platform was integrated into a double-barrel nanoprobe combining pH sensing with feedback-controlled distance dependance via Scanning Ion Conductance Microscopy. This allows for the simultaneous 3D topographical imaging and pHe monitoring of living cancer cells. These classes of nanoprobes were used for real-time high spatiotemporal resolution pHe mapping at the subcellular level and revealed tumour heterogeneity of the peri-cellular environments of melanoma and breast cancer cells.

scholarly journals Nano-Scale Topography Imaging of Live Cell Surface Using Scanning Ion Conductance Microscopy

Hyomen Kagaku ◽  
2015 ◽  
Vol 36 (6) ◽  
pp. 313-318 ◽  
Author(s):  
Hiroki IDA ◽  
Yasufumi TAKAHASHI ◽  
Hitoshi SHIKU ◽  
Tomokazu MATSUE
Keyword(s):  
Cell Surface ◽  
Live Cell ◽  
Ion Conductance ◽  
Scanning Ion Conductance Microscopy ◽  
Nano Scale

scholarly journals Scanning Ion Conductance Microscopy for Studying Biological Samples

Sensors ◽  
2012 ◽  
Vol 12 (11) ◽  
pp. 14983-15008 ◽  
Author(s):  
Patrick Happel ◽  
Denis Thatenhorst ◽  
Irmgard Dietzel
Keyword(s):  
Biological Samples ◽  
Ion Conductance ◽  
Scanning Ion Conductance Microscopy

Imaging Proteins in Membranes of Living Cells by High-Resolution Scanning Ion Conductance Microscopy

2006 ◽  
Vol 45 (14) ◽  
pp. 2212-2216 ◽  
Author(s):  
Andrew I. Shevchuk ◽  
Gregory I. Frolenkov ◽  
Daniel Sánchez ◽  
Peter S. James ◽  
Noah Freedman ◽  
...  
Keyword(s):  
High Resolution ◽  
Living Cells ◽  
Ion Conductance ◽  
Scanning Ion Conductance Microscopy
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